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A System for Microspectrophotometry Employing a Commercial Recording Spectrophotometer

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Abstract

The present paper describes the design and performance of an attachment for the Cary model 14 recording spectrophotometer, which permits the accurate recording of absorption spectra in small areas. A special compartment built into the light path of the spectrophotometer holds a low-magnification microscope (“macroscope”) consisting of two opposed quartz condensers, with which measurements can be made between 300 and 700 mμ in fields 0.1 to 1 mm in diameter. This can be replaced by a conventional microscope with which spectra can be measured from about 350–650 mμ in fields as small as 4 μ in diameter. These arrangements have been used to measure the absorption spectra of visual pigments in situ. With the macroscope, such measurements have been made in small areas of surviving retinas; and with the microscope, they have been made in single isolated outer segments of rods. The present paper contains examples of each type of measurement.

© 1961 Optical Society of America

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